Abstract

Aimof this study is to research an effect of silk fi broin fi lms fabricated by casting method upon Wistar rat’s full-thickness skin wound regeneration.Materials and methods.4 different kinds of fi lms with protein concentration equal to 20 mg/ml were fabricated: fi lms from silk fi broin aqueous solution, fi lms from silk fi broin formic acid solution, fi lms from silk fi broin aqueous solution containing 30% collagen by weight, fi lms from silk fi broin formic acid solution containing 30% collagen by weight. All kinds of fi lms were fabricated by casting method on polished Tefl on surface. Scanning electron microscopy was applied to research fi lms’ surface structure. Cytotoxicity test of the fi lms was realized on mouse 3T3 fi broblasts model by MTT assay. Manufactured fi lms were utilized to regenerate full-thickness skin wounds in Wistar rats.Results.It was shown that fi lms’ surface was characterized by micro- and nanorelief in the form of roughness. The proliferative activity of mouse 3T3 fi broblasts increased during 7 days of cytotoxicity test. Fabricated fi lms enlarge the regeneration rate of full-thickness Wistar rat skin wounds an average of 25%. Histological analysis indicated structural skin restoration without any infl ammatory tissue.Conclusion.All fabricated fi lms are non-cytotoxic and characterized by appropriate structure for the adhesion and proliferation of fi broblasts. The application of fi lms for full-thickness skin wound regeneration increases its restoration rate which is confi rmed by histological examination.

Highlights

  • Aim of this study is to research an effect of silk fibroin films fabricated by casting method upon Wistar rat’s full-thickness skin wound regeneration

  • Cytotoxicity test of the films was realized on mouse 3T3 fibroblasts model by MTT assay

  • The proliferative activity of mouse 3T3 fibroblasts increased during 7 days of cytotoxicity test

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Summary

Получение фиброина шелка

Фиброин шелка получали из коконов тутового шелкопряда B. mori. На первом этапе коконы подвергали очистке от серицина. Навеску шелка из котом XVIII No 3–2016 конов массой 1 г кипятили на водяной бане в 500 мл бидистиллированной воды с добавлением 1260 мг соды в течение 40 минут. Далее кипятили в 500 мл бидистиллированной воды 30 минут и промывали 3,6 л дистиллированной воды. Очищенный фиброин шелка сушили на воздухе при комнатной температуре. Общая концентрация белка в растворе для изготовления пленки составляла 20 мг/мл, содержание коллагена в смеси с фиброином шелка – 30% по массе. Пленки снимали с поверхности тефлона с помощью скальпеля, предварительно инкубировав их в спирте в течение 15 минут, и хранили в 96%-ном спирте при температуре 4 °С

Получение водного раствора фиброина шелка
Анализ структуры пленок методом сканирующей электронной микроскопии
Groups of animals in the experiment of skin regeneration
Моделирование полнослойной кожной раны у крыс
Оценка заживления полнослойной кожной раны
Гистологическое исследование
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